慢性乙型肝炎患者肝组织自然杀伤细胞G2D表达水平的变化及其临床意义

doi:10.3969/j.issn.1672-5069.2019.03.008

摘要/Abstract

摘要： 目的探讨慢性乙型肝炎病毒（HBV）感染者自然杀伤细胞（NK）G2D表达水平及临床意义。方法 2015年7月~2018年7月我院收治的慢性HBV携带者20例、慢性乙型肝炎（CHB）患者18例和HBV相关慢加急（亚急性）肝衰竭患者16例,另选健康体检者20例作为对照,患者接受肝活检,正常肝组织取自肝血管瘤患者手术获得的肝组织。使用流式细胞术检测单个核细胞中NK细胞和NKG2D⁺NK细胞百分比,采用ELISA法检测血清穿孔素和颗粒酶B水平,采用荧光定量PCR法检测肝组织NKG2D mRNA,采用Western blot法检测NKG2D蛋白表达水平。组间比较采用单因素方差分析,两两比较采用SNK法。结果 HBV携带者NK细胞频率和NKG2D⁺NK细胞频率分别为（5.1±1.5）%和（3.4±1.1）%,CHB患者分别为（8.5±2.0）%和（12.3±2.1）%,肝衰竭患者分别为（8.8±1.6）%和（19.0±2.4）%,健康人分别为（14.2±2.1）%和（7.5±1.5）%,差异显著（P<0.05）;HBV携带者血清穿孔素和颗粒酶B水平分别为（15.2±3.2）ng/L和（23.0±3.5）ng/L,CHB为（31.5±3.6）ng/L和（52.5±4.0）ng/L,肝衰竭为（60.2±11.0）ng/L和（126.8±30.1）ng/L,健康人为（13.9±4.0）ng/L和（21.1±3.9）ng/L,差异显著（P<0.05）;HBV携带者肝组织NKG2D mRNA和NKG2D蛋白水平分别为（0.1±0.0）和（0.0±0.0）,CHB为（3.5±0.9）和（1.1±0.2）,肝衰竭为（7.0±1.2）和（1.8±0.2）,健康肝组织为（0.6±0.2）和（0.3±0.0）,差异显著（P<0.05）。结论 NKG2D可能通过NK细胞合成分泌穿孔素和颗粒酶B等杀伤介质因子发挥效应。不同免疫状态下慢性HBV感染者NKG2D水平存在差异,下调NKG2D表达可能通过抑制NK细胞而改善肝组织免疫损伤。

关键词: 乙型肝炎, 慢加急（亚急性）肝衰竭, 自然杀伤细胞G2D

Abstract: Objectiv The aim of this study was to investigate the G2D expression of peripheral blood natural killer cells in patients with chronic hepatitis B（CHB）. Methods 20 chronic HBV carriers,18 patients with CHB,16 patients with HBV-induced acute-on-chronic liver failure and 20 healthy individuals were recruited in our hospital between July 2015 and July 2018. The liver biopsies were performed in all patents and normal liver tissue was obtained from the adjacent liver tissues of patients with hepatic hemangioma. Flow cytometry was used to detect NK cells and NKG2D⁺NK cells in peripheral blood,serum perforin and granulase B levels were detected by ELISA,and liver tissue NKG2D mRNA and NKG2D protein expression were detected by fluorescence quantitative PCR and Western bloting,respectively. Results The frequencies of NK cells and NKG2D⁺NK cells in HBV carriers were （5.1±1.5）% and （3.4±1.1）%,in patients with CHB were （8.5±2.0）% and （12.3±2.1）%,in patients with liver failure were （8.8±1.6）% and （19.0±2.4）%,and control were （14.2±2.1）% and （7.5±1.5）% （P<0.05）;serum levels of perforin and granzyme B were （15.2±3.2） ng/L and （23.0±3.5） ng/L,（31.5±3.6） ng/L and （52.5±4.0） ng/L,（60.2±11.0） ng/L and （126.8±30.1） ng/L,and （13.9±4.0） ng/L and （21.1±3.9） ng/L） in the four groups（P<0.05）;the NKG2D mRNA levels and NKG2D protein expression were（0.1±0.0） and （0.0±0.0）,（3.5±0.9） and（1.1±0.2）,（7.0±1.2） and （1.8±0.2）,and （0.6±0.2） and （0.3±0.0） in the four groups （P<0.05）. Conclusion NKG2D might exert its action through the synthesis and secretion of perforin and granulase B by NK cells,and down-regulation of NKG2D might improve liver injury and prevent liver failure.

Key words: Hepatitis B, Acute-on-chronic liver failure, Natural killer cells, NKG2D

钟宋, 刘玉元, 刘树人, 周国柱, 刘崇文, 欧阳石. 慢性乙型肝炎患者肝组织自然杀伤细胞G2D表达水平的变化及其临床意义[J]. 实用肝脏病杂志, 2019, 22(3): 341-344.

Zhong Song, Liu Yuyuan, Liu Shuren. G2D expression of peripheral blood natural killer cells in patients with chronic hepatitis B[J]. JOURNAL OF PRACTICAL HEPATOLOGY, 2019, 22(3): 341-344.

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